In the fields of molecular biology, biochemistry and pharmacology an accurate and expeditious analysis of biomolecules such as recombinant deoxyribonucleic acid (DNA) is of the utmost importance: Typically, a DNA specimen is analyzed by placing the specimen into a porous gel matrix, which allows the movement of particles but impedes the rate of travel. A current is then applied to the gel matrix to produce positively and negatively charged ends of the gel matrix. Under these conditions the DNA migrates toward the positively charged end of the gel matrix. This process is used to separate DNA of different sizes and to separate newly synthesized DNA strands with labels in order to elucidate sequences of small DNA strands.
Unfortunately, the above-described process is inadequate for the sequencing of long DNA strands. DNA longer than 10 to 20 thousand bases cannot be reliably synthesized in a single polymerization reaction. To provide sequence data, the DNA synthesized in a reaction must start from the same position on the DNA strand being sequenced so that an exact base at which the reaction terminated with a label is known. If the reaction is started randomly, DNA fragments would represent all possible bases within a sample, but the DNA would be separated only by size. The result would be fragments of all possible sizes terminated with all possible bases. Even if the entire sequence of each fragment were known, the task of overlapping the fragments into a complete genome would be a difficult and error prone. Roughly forty-percent of the human genome is composed of non-functional copies of viral genes and the overlapping regions are similar throughout. The result is that some regions will be placed out of order and others will likely be omitted because they are identical to other fragments. To provide data that is based on more than the length of the DNA fragment and the terminating base of the fragment, there must be a process of controlling the orientation and the movement of the DNA to be examined.
A cyclotron is a particle accelerator device that is still used in hospitals to produce activated technetium and other isotopes that have short half lives. The device is simply a track that runs in a circular path with plates capable of carrying a charge when switches are closed. The plates are charged so that electrons are drawn one way and repelled from the opposite direction. The switches are closed at an increasing rate, which causes a particle to be repelled from one side and pulled from the other side along the track at an increasing velocity until a collision is desired. A door in the track is then opened and the accelerating electrons are ejected directly toward the substance to be bombarded with beta particles.
This principle inherent in the cyclotron could theoretically direct the movement of DNA alone, however the fragments of DNA would still be tightly condensed and no information other than the position of the DNA would be known. To remedy this problem, a linear track is placed along a linear path, and rather than charged plates, an electric field is applied such that charges of one polarity are pulled across the track, and charges having the opposite polarity are repelled. Alternatively, the field could be designed to attract a magnetic particle while repelling a charged particle. The DNA would then be pulled or pushed as an indecipherable mass. To decipher the mass, it is necessary to add another component such as an antibody. Antibodies specific to the 5′ or 3′ end of a DNA strand can be generated and attached to additional particles, including charged or magnetic particles.
If the 5′ end of a DNA strand is attached by an antibody to a charged bead that is pulled down the linear track, and the 3′ end is attached to a bead that is repelled less strongly than the 5′ end is attracted, there will be a net movement along the linear track. Thus, the direction and the order of a DNA sample can be controlled. The only limitation to the length of a DNA fragment that can be moved is the length of the linear track. A linear track in conjunction with antibody bound charged beads requires a sensor that is capable of making direct observations of DNA strands.
The inventive biomolecule analyzing system that utilizes a ciliated sensor solves the problems inherent in the prior art.
A search of the prior art did not disclose any patents that read directly on the claims of the instant invention. However the following U.S. patents are considered related:
PATENT NO.INVENTORISSUED7,129,047Yamashita31 Oct. 20066,924,105Sudo, et al2 Aug. 20056,670,131Hashimoto30 Dec. 20036,573,089Vann3 Jun. 2003
The U.S. Pat. No. 7,129,047 discloses a nucleotide detector that consists of metal particles and single-stranded thiol DNAs. The metal particles are placed on the surface of a substrate. The DNAs have sulphur atoms at their ends, which are bonded to gold particles and placed uniformly over the substrate. Therefore, once a fluorescence-labeled single-stranded DNA is hybridized with any of the thiol DNAs, a high fluorescence intensity is stably obtained. The nucleotide detector is therefore usable as a high-performance DNA sensor with a high SN ratio.
The U.S. Pat. No. 6,924,105 discloses a method for directly analyzing double-stranded DNA that is present in an analyte without degeneration. The method comprises the steps of:
(1) contacting the analyte with a double-stranded DNA recognizing substance immobilized on a support, and
(2) measuring double stranded DNA that are bound to the double stranded DNA recognizing substance.
The U.S. Pat. No. 6,670,131 discloses a nucleic acid detection apparatus. The apparatus includes a nucleic acid immobilized electrode, a plurality of vessels for bringing the nucleic acid probe into contact with a subject substance, a counter electrode disposed on a bottom surface or inside surface of the vessels, and an electric circuit for applying a voltage between the nucleic acid immobilized electrode and the counter electrode. A nucleic acid is detected by inserting the nucleic acid immobilized electrode into each vessel containing the subject substance, and using the counter electrode disposed on the bottom surface or inside surface of the vessel to electrically control a reaction.
The U.S. Pat. No. 6,573,089 discloses an apparatus and method for contacting at least two chemical species: The apparatus comprises a support plate having a channel for receiving a mobile chemical species and a fiber having a second immobilized chemical specie disposed on the support plate. A portion of the fiber is exposed to the channel such that the mobile chemical species is capable of contacting the second chemical species.
For background purposes and as indicative of the art to which the invention is related reference my be made to the remaining patents located in the patent search:
PATENT NO.INVENTORISSUED7,123,029Frey, et al17 Oct. 20066,916,614Takenaka12 Jul. 20056,905,829Cho, et al14 Jun. 20056,893,824Ito17 May 20056,890,764Chee, et al10 May 20056,812,005Fan, et al2 Nov. 20046,667,159Walt, et al23 Dec. 20036,649,404Vann, et al18 Nov. 20036,620,584Chee, et al16 Sep. 2003